Physiologically active substance derived from malt and process for the production thereof

ABSTRACT

A multi-functional physiologically active substance can be obtained from the residue of saccharified malt which is worthless other than feed for cattle. The isolation of the active substance is performed by an extraction of the residue with water or an aqueous solvent and removal of a low molecular weight fraction from the extract.

This is a division, of application Ser. No. 539,850, filed Jun. 18,1990, now abandoned.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to a physiologically active substance extractedfrom the residue of saccharified malt and the process for the productionthereof. "Physiologically active " means, for example, the inhibition ofplatelets aggregation, prevention of death due to acute pulmonaryembolism, improvement of erythrocyte transformation, analgesic activity,anti-nephropathic action, radiation exposure protection activity,hypotensive activity, and hypoglycemic activity.

2. Description of the Prior Art

Few efforts have been known to extract and produce a useful substancefrom the residue of saccharified malt. For example, Japanese Laid-OpenApplication No. 129776 (1976) describes an extraction of a solidnutritious protein from brewed cereals such as residue of maltsaccharification. However, no attempts have been known to obtain aphysiologically active polymer from the waste of saccharified malt.

SUMMARY OF THE INVENTION

The present inventors have been searching for substances useful for themaintenance of human health from unused resources employed forindustrial purposes. Unexpectedly, a physiologically active polymerwhich exhibits various useful properties for the maintenance of humanhealth including inhibition of platelet aggregation and improvement inthe transformation of erythrocytes was isolated by an extraction ofresidue of malt saccharification with water and accomplished the presentinvention. Therefore, the main purpose of the present invention is toprovide a physiologically active substance from the filtered residue ofa saccharified malt, by an extraction with water or an aqueous solvent,and removal of a low molecular weight fraction, and the procedure forthe production thereof. Other purposes of the present invention willbecome apparent by the following description.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a chromatogram of the dried extract explained in example 1.

FIG. 2, 3 and 4 show an infrared absorption spectrum, a nuclear magneticresonance spectrum and ultraviolet and visible absorption spectrum ofthe dried extract, respectively.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The physiologically active substance of the present invention obtainedfrom the residue of saccharified malt can be defined by the followingproperties:

(i) Molecular weight of approximately 2,000 or more.

(ii) Infrared absorption (IR) spectrum having peaks at 3,600-3,200 cm⁻¹and 1,700-1,600 cm⁻¹.

(iii) Proton nuclear magnetic resonance (NMR) spectrum having peaks at0.5-6.0 ppm and 6.0-8.5 ppm.

(iv) Ultraviolet absorption (UV) spectrum having peaks at 200-250 nm and250-350 nm.

(v) Positive to the phenol sulfuric acid color reaction.

(vi) Positive to the copper-Folin color reaction.

The substance of the present invention can be produced by the followingprocedure.

Extraction of a filtered residue prepared by saccharification of maltwith water or an aqueous solvent, and removal of a low molecular weightfraction to isolate the desired fraction.

The malt of the present invention is prepared by germinating the grainsincluding gramineous crops. These grains include e.g. barley, wheat,rye, oats, naked barley and corn. The grains are pulverized andsaccharified in warm water, and starches may be added if necessary. Thesaccharified mixture is filtered and the residue or filter cake is usedfor the raw material of the present invention. The residue has been usedfor cattle feed and the filtrate or wort is used for the production ofbeer.

The above mentioned filtered residue is then extracted with water or anaqueous solvent. Water includes preferably hot, warm and 0.02-2.0N warmalkaline aqueous solution and the aqueous solvents include hydrophilicorganic solvents such as methanol, ethanol and acetone or their aqueousmixture. The extraction is performed by the addition of 10-1,000,000weight parts of water or the aqueous solvent to 100 weight parts ofdried or wet filter cake, and for a few hours to overnight. Stirring,ultrasonication or an addition of a surface active agent may be used forefficient extraction. The extraction is performed generally attemperatures of 10°-150° C., preferably at 40°-120° C., and morepreferably at 60°-100° C. When an alkaline solution is used for theextraction, the resulted extract is neutralized and the formedprecipitates are filtered, then a low molecular weight fraction of lessthan 2,000 is separated from the obtained supernatant. The extractionwith an 0.1-1.0N alkaline solution provides superior extraction ratewith higher water solubility of the extract than that with water withoutaffecting the properties of extract. The extract then may be made pH 4.0to precipitate a protein. The obtained supernatant is furtherneutralized to pH 7.0 to remove precipitates. A low molecular weightfraction in the supernatant is removed by various methods such asdialysis, salting out, ultrafiltration, reverse osmosis, gel filtration,or precipitation by the addition of an organic solvent to give thephysiologically active fraction of the present invention. The obtainedfraction can be used as it is, or may be processed before use such ascondensation, lyophilization and spray drying.

The substance or extract of the present invention has the followingproperties:

(i) Molecular weight of approximately 2,000 or more.

(ii) Infrared absorption spectrum having peaks at 3,600-3,200 cm⁻¹ and1,700-1,600 cm⁻¹.

(iii) Proton nuclear magnetic resonance spectrum having peaks at 0.5-6.0ppm and 6.0-8.5 ppm.

(iv) Ultraviolet absorption spectrum having peaks at 200-250 nm and250-350 nm.

(v) Positive to the phenol sulfuric acid color reaction.

(vi) Positive to the copper-Folin color reaction.

The physiologically active substance of the present invention showed noacute toxicity in oral administration in mice as shown in the followingExample 3. The physiological activities of the substance wereinvestigated and confirmed by the following examples of 4-12 exhibitingsuch as the inhibition of platelets aggregation, prevention of death dueto acute pulmonary embolism, improvement of erythrocyte transformation,analgesic activity, anti-nephropathic activity, radiation exposureprotecting activity, hypotensive activity, and hypoglycemic activity.These properties are particularly useful for the geriatric diseases ofvarious systems such as central nervous system, circulatory system,urinary system, an metabolic system which are expected to become moreimportant in the future. The substance is safe and very useful for themaintenance of human health and will be one of the awaited medicines.

The physiologically active substance of the present invention can beused as an anti-thrombotic agent, anti-ischemic agent, analgesic,anti-nephrophathic agent, radiation exposure protecting agent,anti-hypertensive agent, and anti-diabetic agent for the prevention andtreatment of above mentioned diseases in the forms of medicines or foodsand drinks. The substance can be orally administered at 0.01-10,000mg/kg daily in several divided forms such as tablets, powders, granules,or capsules. The substance can be used as foods and drinks and can betaken at 0.01-10,000 mg/kg daily as it is or as an additive for theprocessed cakes such as biscuits, crackers, snacks, rice cakes,chocolates, cocoas, chewing gums, candies, western, Japanese and Chinesecakes, doughnuts, pizzas, crepes, chilled desserts, breads and buns, andnoodles, macaronies, pickles, tofu (bean curds), processed meats such ashams and sausages, milk products such as butters, cheeses, yogurts, milkand ice creams, and margarines, hamburgers and seasonings. The substancecan be added to drinks such as carbonated water, soda pops, ciders,fruits juices, pulverized fruit juices, mineral water, soya milk,coffer, tea, green tea, toasted tea, and oolong tea.

The present invention is further explained by the following examples,but these examples are illustrated to exemplify the present inventionand the scope of the present invention is not restricted by theseexamples.

EXAMPLE 1

Ten kg of barley were germinated, mixed with proper amount of starch andsaccharified by a conventional method. The saccharified mixture wasfiltered and the filtered residue was dried.

One hundred gram of the saccharified residue was added to four L of 0.5NNaOH and extracted for three hours under boiling. The extract was madepH 4.0 with 1N HCl and allowed to stand. The formed precipitates wereseparated by centrifugation at 6,000 rpm for 30 minutes. The obtainedsupernatant was neutralized to pH 7.0 with 1N NaOH and centrifuged againat 6,000 rpm for 30 minutes. The obtained supernatant was condensedunder reduced pressure, dialyzed against running water (Viskingcellulose) and lyophilized to give 23 g of the dried extract.

The obtained dried extract showed weight-average molecular weight of209,000 by GPC-Lalls method. One example of a gel chromatogram of thedried extract is shown in FIG. 1 [packing material: CellulofineGLC-2000-C, column: φ 2.5 cm×height 45 cm, fractionation: three ml/onefraction, detection method: color reaction with phenol sulfuric acid(490 nm), sample: injection by 30 mg/3 ml]. FIG. 1 shows the separationof a fraction of molecular weight of 10,000 or less. FIG. 2 shows an IRspectrum of the substance determined with KBr tablet by the IRspectrophotometer type A202 [Japan Spectroscopic Co., Ltd. (JASCO)].Absorption peaks at 3,600-3,200 cm⁻¹ and 1,700-1,600 cm⁻¹ were observed.FIG. 3 shows a proton nuclear magnetic resonance (PMR) spectrum wasdetermined with an internal standard of DSS by JMN-GSX type 500spectrometer [Nihon Denshi Ltd. (JEOL)] at 500 MHz, revealing peaks at0.5-6.0 ppm and 6.0-8.5 ppm. FIG. 4 shows UV and visible absorptionspectra of the fraction with peaks at 2090-250 nm and 250-350 nmdetermined by Multipurpose Recording Spectrophotometer MPS-2000(Shimadzu Seisakusho Ltd.). These Fig. of chromatogram and spectrarepresent examples of the properties of the dried extract of the presentinvention, and the scope of dried extract of the present invention isnot defined by these Fig.

The dried extract showed positive to phenol sulfuric acid color reactionrevealing the presence of sugars. The composition of the sugars wereanalyzed after acidic hydrolysis by liquid chromatography LC-6A(Shimadzu Seisakusho Ltd.) and sugars such as arabinose, xylose, glucoseand galactose were detected and confirmed. The copper-Folin colorreaction was also positive indicating the presence of a protein. Thecontained amino acids in the protein were analyzed after acid hydrolysisby Hitachi 835 (Hitachi Ltd.) amino acid analyzer and common acidicamino acids, basic amino acids and hydrophobic amino acids were detectedand confirmed.

EXAMPLE 2

Malts of wheat, rye, oats and naked barley were treated similar mannerto Example 1 and resulted dried extracts, respectively. Thephysicochemical properties of the dried extracts were similar to thoseof extract of barley malt as shown in Table 1.

                  TABLE 1                                                         ______________________________________                                                  Resource of extract of malt residue                                                             Naked                                             Item        Barley  Wheat   barley Rye   Oats                                 ______________________________________                                        Average M.W.                                                                              20.9    20.5    22.1   21.8  19.8                                 (10,000)                                                                      Solubility  Freely  Freely  Freely Freely                                                                              Freely                               in water    soluble soluble soluble                                                                              soluble                                                                             soluble                              Qualitative posi-   posi-   posi-  posi- posi-                                Test of sugars                                                                            tive    tive    tive   tive  tive                                 (phenol sulfu-                                                                ric acid                                                                      method)                                                                       Qualitative posi-   posi-   posi-  posi- posi-                                test of pep-                                                                              tive    tive    tive   tive  tive                                 tides (Copper-                                                                Folin method)                                                                 IR spectra                                                                    3,600-3,200 cm.sup.-1                                                                     posi-   posi-   posi-  posi- posi-                                1,700-1,600 cm.sup.-1                                                                     tive    tive    tive   tive  tive                                 NMR spectra                                                                   0.5-6.0 ppm posi-   posi-   posi-  posi- posi-                                6.0-8.5 ppm tive    tive    tive   tive  tive                                 UV absorption                                                                 spectra                                                                       200-250 nm  posi-   posi-   posi-  posi- posi-                                250-350 nm  tive    tive    tive   tive  tive                                 ______________________________________                                    

EXAMPLE 3

The safety of the extracts of the present invention derived of malts wastested by the method shown below.

Each of ten ICR mice (male, five weeks old) were enrolled in the study.Each of the dried extracts obtained by Examples 1 and 2, 5 g, weredissolved in 10 ml of distilled water and orally administered by gavageat five g/kg and observed for 14 days. No dead mouse was found. The micewere sacrificed and their blood were drawn. No abnormal result was foundin general blood test, blood biochemistry an pathological tests. Anexcellent safety of the substance was found in those acute toxicitytests.

The following examples 4-12 show the physiological activities of thedried extracts derived from malt obtained by the present invention.

EXAMPLE 4

Fresh blood was drawn by citrate added syringe from nine weeks oldfemale Donryu rats according to the method of G. V. R. Born [Nature,194, 927 1962] and PRP (platelet rich plasma) was obtained bycentrifugation. Dried extracts obtained according to Examples 1 and 2were added, respectively, to the PRP at 2.5 mg/ml and 50 μM ofadenosine-5'-diphosphate (ADP) was added as a platelet aggregationstimulant to investigate the inhibitory activity on plateletaggregation. The control group used PRP without containing the extract.

The determination of platelet aggregation was performed by measuring thetransparency of PRP using an aggrecometer (Biodata Corp.). Theinhibitory rate, I, of the extract was calculated by the followingequation. ##EQU1##

The results are shown in Table 2, revealing the excellent inhibitoryactivity on platelet aggregation. The results also indicate that theextract of the present invention can be appllied for the prevention andtreatment of thrombus formation due to its inhibitory activity, I,against platelet aggregation.

                  TABLE 2                                                         ______________________________________                                        Resource of   Inhibitory rate of                                              extract of malt                                                                             platelet aggregation,                                           residue       I                                                               ______________________________________                                        Barley        ++                                                              Wheat         +                                                               Naked barley  +                                                               Rye           ++                                                              Oats          ++                                                              ______________________________________                                    

The inhibitory rates, I, in the above Table 2 are classified by thefollowing grades:

    ______________________________________                                        -;           0       ≦ I <                                                                          20(%)                                            +;           20(%)   ≦ I <                                                                          30(%)                                            ++;          30(%)   ≦ I ≦                                                                   100(%)                                           ______________________________________                                    

EXAMPLE 5

On thousand mg of the dried extract obtained by Example 1 or 2 wasdissolved in 20 ml of distilled water and orally administered by gavageto four weeks old male ICR mice according to the method of Narday et al.at a rate of 1,000 mg/kg. After three hours, 400 mg/kg of ADP wasadministered to the mice in tail vein in three seconds and the mice wereobserved for 10 minutes to obtain the death rate for the investigationof the preventive effect against death due to acute pulmonary embolism.The control group was administered 20 ml/kg of distilled water insteadof the extract. The results are shown in Table 3 revealing an excellentpreventive effects against death due to acute pulmonary embolism.

The results also indicate that the extract of the present invention canbe applied for the prevention and treatment of thrombus formation due toits preventive activity against death caused by acute pulmonaryembolism, I.

                  TABLE 3                                                         ______________________________________                                        Resource of   Inhibitory rate of death                                        extract of malt                                                                             due to acute pulmonary                                          residue       embolism, I                                                     ______________________________________                                        Barley        ++                                                              Wheat         +                                                               Naked barley  +                                                               Rye           +                                                               Oats          ++                                                              ______________________________________                                    

The inhibitory rates, I, in the above Table 3 are classified by thefollowing grades:

    ______________________________________                                        -;           0       ≦ I <                                                                          20(%)                                            +;           10(%)   ≦ I <                                                                          30(%)                                            ++;          30(%)   ≦ I ≦                                                                   100(%)                                           ______________________________________                                    

The inhibitory rate of death due to acute pulmonary embolism, I, iscalculated by the following equation. ##EQU2##

EXAMPLE 6

One thousand mg of the dried extract obtained by Example 1 or 2 wasdissolved in 20 ml of distilled water and orally administered by gavageto eight weeks old female Donryu rats according to the method of Furlowet al. [Science. 187, 658 (1975)] at a rate of 1,000 mg/kg. After 30minutes, 20 mg/kg of arachidonic acid was administered to the ratsintra-arterially directing toward the brain. The rats were observed toobtain the death rate for the investigation of the preventive effectagainst death due to thrombus. The control group was administered 20ml/kg of distilled water instead of the extract. The results are shownin Table 4 revealing an excellent preventive effects against death dueto thrombus.

The results also indicate that the extract of the present invention canbe applied for the prevention and treatment of ischemia due to itspreventive activity against death caused by thrombus, I.

                  TABLE 4                                                         ______________________________________                                        Resource of   Inhibitory rate of death                                        extract of malt                                                                             due to thrombus,                                                residue       I                                                               ______________________________________                                        Barley        +                                                               Wheat         ++                                                              Naked barley  +                                                               Rye           +                                                               Oats          +                                                               ______________________________________                                    

The inhibitory rates, I, in the above Table 4 are classified by thefollowing grades:

    ______________________________________                                        -;           0       ≦ I <                                                                          10(%)                                            +;           10(%)   ≦ I <                                                                          30(%)                                            ++;          30(%)   ≦ I ≦                                                                   100(%)                                           ______________________________________                                    

The inhibitory rate of death, I, due to thrombus is calculated by thefollowing equation. ##EQU3##

EXAMPLE 7

One thousand mg of the dried extract obtained by Example 1 or 2 wasdissolved in one ml of distilled water and orally administered by gavageto 10 weeks old female Donryu rats according to the method of Otomatsuet al. at a rate of 1,000 mg/kg. After three hours, the blood was drawnfrom ventral aorta and added to 10 fold physiological saline solution.The mixture was stirred by Termo-mixer and hemolyzed and centrifugatedat 1,500 rpm for five minutes. The content of hemoglobin in the obtainedsupernatant was determined by an absorption 540 nm.

The rate of hemolysis was calculated on the base hemolysis rate of 100obtained by the addition of distilled water and investigated theimprovement of transformation of erythrocytes by the substance of thepresent invention. The control group was administered one ml/kg ofdistilled water instead of the extract.

The results are shown in Table 5 revealing an excellent effect ofimprovement in the transformation of erythrocytes. The results alsoindicate that the extract of the present invention can be applied forthe prevention and treatment of ischemia due to its activity on theimprovement of transformation of erythrocytes, P.

                  TABLE 5                                                         ______________________________________                                        Resource of   Improved rate on the                                            extract of malt                                                                             transformation of                                               residue       erythrocyte, P                                                  ______________________________________                                        Barley        ++                                                              Wheat         +                                                               Naked barley  ++                                                              Rye           ++                                                              Oats          +                                                               ______________________________________                                    

The improved rates, P, in the above Table 5 are classified by thefollowing grades:

    ______________________________________                                        -;           0       ≦ P <                                                                          10(%)                                            +;           10(%)   ≦ P <                                                                          30(%)                                            ++;          30(%)   ≦ P ≦                                                                   100(%)                                           ______________________________________                                    

The improved rate of erythrocyte transformation, P, is calculated by thefollowing equation. ##EQU4##

EXAMPLE 8

One thousand mg of the dried extract obtained by Example 1 or 2 wasdissolved in 10 ml of distilled water and orally administered by gavageto six weeks old female ICR mice according to the method described inFundamental Course of Medicine, vol. 6, Evaluation of Drug Efficacy (1)Pharmacological Tests (Part 1) p. 283. Antipyretics Analgesic, Pub. byChijin Shokan (1971), at a rate of 1,000 mg/kg. After 30 minutes, 0.1ml/10 g of body weight of 2.0% acetic acid solution wasintraperitoneally administered. After 10 minutes, the number of writhingwas counted for 10 minutes to evaluate the analgesic effect of theextract. The control group was administered 10 ml/kg of distilled waterinstead of the extract.

The results are shown in Table 6 revealing an excellent analgesic effectof the substance of the present invention. The results also indicatethat the extract can be applied for the diseases and symptoms whichrequire the relief of pain.

                  TABLE 6                                                         ______________________________________                                        Resource of                                                                   extract of malt                                                                              Analgesic effect,                                              residue        I                                                              ______________________________________                                        Barley         +                                                              Wheat          +                                                              Naked barley   +                                                              Rye            +                                                              Oats           ++                                                             ______________________________________                                    

The rates of activity, I, in the above Table 6 are classified by thefollowing grades:

    ______________________________________                                        -;           0       ≦ I <                                                                          10(%)                                            +;           10(%)   ≦ I <                                                                          30(%)                                            ++;          30(%)   ≦ I ≦                                                                   100(%)                                           ______________________________________                                    

The rate of analgesic effect, I, is calculated by the followingequation. ##EQU5##

EXAMPLE 9

Eight weeks old female Wistar rats were subcutaneously administered 10mg/kg of an aminonucleoside for consecutive days to induce nephrosis.One thousand mg of the dried extract obtained by Example 1 or 2 wasdissolved in 20 ml of distilled water and orally administered by gavagefrom five days before the administration of the aminonucleoside forconsecutive 14 days according to the method of Caulfied et al. Urinaryprotein on the final day of the extract administration was determined toinvestigate the anti-nephrotic effect. The control group wasadministered 20 ml/kg of distilled water instead of the extract.

The results are shown in Table 7 revealing an excellent decrease ofurinary protein concentration. The results also indicate that theextract can be widely applied for the prevention and treatment ofnephropathy.

                  TABLE 7                                                         ______________________________________                                        Resource of    Inhibitory rate of                                             extract of malt                                                                              urinary protein,                                               residue        I                                                              ______________________________________                                        Barley         ++                                                             Wheat          ++                                                             Naked barley   +                                                              Rye            ++                                                             Oats           ++                                                             ______________________________________                                    

The inhibitroy rates, I, in the above Table 7 are classified by thefollowing grades:

    ______________________________________                                        -;           0       ≦ I <                                                                          10(%)                                            +;           10(%)   ≦ I <                                                                          30(%)                                            ++;          30(%)   ≦ I ≦                                                                   100(%)                                           ______________________________________                                    

The inhibitory rate of urinary protein, I, is calculated by thefollowing equation. ##EQU6##

EXAMPLE 10

Eight weeks old male C3H/HeJ mice were systematically irradiated (3 Gy)under non-restraint condition. One thousand mg of the dried extractobtained by Example 1 or 2 was dissolved in 10 ml of distilled water andorally administered from the next day of the irradiation for consecutivedays. After 12th day of the irradiation, blood was drawn from ophthalmicvein under ether anesthesia and leukocyte count which is often used forthe index of the influence of radiation exposure was determined and theinhibitory rate of leukopenia, I, was calculated by the followingequation: ##EQU7##

The control group was administered 10 ml/kg of distilled water insteadof the extract.

The results are shown in Table 8 revealing an inhibitory effect of thesubstance of the present invention against leukopenia caused byradiation exposure. The results also indicate a possibility of theextract for the application to the prevention and treatment of thedecline of vital functions due to the radiation exposure and the extractwill become a very important substance for terrestrial livings includinghuman being under the progress of radioactive contamination.

                  TABLE 8                                                         ______________________________________                                        Resource of    Inhibitory rate of                                             extract of malt                                                                              leukopenia,                                                    residue        I                                                              ______________________________________                                        Barley         ++                                                             Wheat          +                                                              Naked barley   +                                                              Rye            ++                                                             Oats           +                                                              ______________________________________                                    

The inhibitory rates, I, in the above Table 8 are classified by thefollowing grades:

    ______________________________________                                        -;           0       ≦ I <                                                                          10(%)                                            +;           10(%)   ≦ I <                                                                          20(%)                                            ++;          20(%)   ≦ I ≦                                                                   100(%)                                           ______________________________________                                    

EXAMPLE 11

One thousand mg of the dried extracts obtained by Examples 1 or 2 wasdissolved in 20 ml of distilled water and orally administered by gavageat a rate of 1,000 mg/kg to 16 weeks old male spontaneously hypertensiverats (hereinafter abbreviated as SHR rats). After three hours, the bloodpressure were determined to examine the hypotensive effect. The bloodpressure was determined by Rat Tail Manometer-Touchmeter System KN-209(Natsume Seisakusho Co., Ltd.) after keeping the SHR rats in a pre-heatbox (Natsume Seisakusho Co., Ltd.) at 45° C. for two minutes. Thecontrol group was administered 20 ml/kg of distilled water instead ofthe extract. The results are shown in Table 9, revealing an excellenthypotensive effect of the substance in SHR rats. The results alsoindicate the possibility of the application of extract for theprevention and treatment of hypertensions.

                  TABLE 9                                                         ______________________________________                                        Resource of                                                                   extract of malt                                                                              Hypotensive rate,                                              residue        L                                                              ______________________________________                                        Barley         ++                                                             Wheat          +                                                              Naked barley   +                                                              Rye            ++                                                             Oats           ++                                                             ______________________________________                                    

The hypotensive rates, L, in the above Table 9 are classified by thefollowing grades:

    ______________________________________                                        -;           0       ≦ L <                                                                          10(%)                                            +;           10(%)   ≦ L <                                                                          20(%)                                            ++;          20(%)   ≦ L ≦                                                                   100(%)                                           ______________________________________                                    

The hypotensive rate, L, is calculated by the following equation.##EQU8##

EXAMPLE 12

Sixty mg/kg of streptozocin was intravenously administered once to 10weeks old male Wistar rats according to the method of Hatanaka et al.Five days after the administration, urine sugar and blood sugar weredetermined. Rats confirmed with the diabetes and hyperglycemia wereenrolled the test. One thousand mg of the dried extract obtained byExample 1 of 2 was dissolved in 20 ml of distilled water andadministered orally by gavage at a rate of 1,000 mg/kg to the enrolledrats. After three hours, the blood was drawn and blood sugar wasdetermined by RABA Super. The control group was administered 20 ml/kg ofdistilled water instead of the extract. The results are shown in Table10, revealing an excellent hypoglycemic effect of the extract of thepresent invention in streptozocin induced diabetes model rats. Theresults also indicate that the extract can be applied for the preventionand treatment of diabetes.

                  TABLE 10                                                        ______________________________________                                        Resource of                                                                   extract of malt                                                                             Hypoglycemic rate,                                              residue       L                                                               ______________________________________                                        Barley        ++                                                              Wheat         +                                                               Naked barley  +                                                               Rye           +                                                               Oats          ++                                                              ______________________________________                                    

The hypoglycemic rates, L, in the above Table 10 are classified by thefollowing grades:

    ______________________________________                                        -;           0       ≦ L <                                                                          10(%)                                            +;           10(%)   ≦ L <                                                                          15(%)                                            ++;          15(%)   ≦ L ≦                                                                   100(%)                                           ______________________________________                                    

The hypoglycemic rate, L, is calculated by the following equation.##EQU9##

EXAMPLE 13

This Example exemplifies a pharmaceutical preparation of the extract ofthe present invention.

One kg of the spray dried extract obtained by Example 1 or 2 was mixedwith one kg of lactose, 500 g of hydroxypropyl-cellulose and 20 g ofcalcium stearate. The mixture was prepared tablets having a diameter of8 mm and weighing 252 mg/tablet by a rotary tabletting machine (HT-9type).

The following Examples 14-19 illustrate the practical applications ofthe extract of the present invention to foods and drinks.

EXAMPLE 14

Dried extract obtained by Example 1 or 2 was sieved and sterilized.Three g of the sterilized product and 58 g of commercial powdered drinkwere mixed in a mortar for 30 minutes to give a powdered drinkcontaining the extract of the present invention.

EXAMPLE 15

Three parts of the dried extract obtained by Example 1 or 2 was added toa mixture of 100 parts of wheat flour, 1.4 parts of salt, 0.14 part ofbrine (dried weight, seasoning for cooking), 32 parts of water, and asmall amount of starch and natural rubber. The obtained mixture waskneaded, rolled, formed to noodles, steam heated, cut, molded, fried andcooled to give a fried noodles.

EXAMPLE 16

One hundred parts of a commercial orange juice and two parts of thedried extract obtained by Example 1 or 2 were thoroughly mixed to givean orange drink containing the extract of the present invention.

EXAMPLE 17

Five parts of the dried extract obtained by Example 1 or 2 and 100 partsof a commerical milk were mixed thoroughly to give a milk containing theextract of the present invention.

EXAMPLE 18

One part of the dried extract obtained by Example 1 or 2 and 100 partsof commercial mineral water were mixed thoroughly to give a mineralwater containing the extract of the present invention.

EXAMPLE 19

Ten parts of the dried extract obtained by Example 1 or 2 and 100 partsof a commercial hot cake mix powder were mixed thoroughly to give a hotcake mix (powder form) containing the extract of the present invention.The obtained product can be used to prepare delicious hot cakes similarto a conventional hot cake mix (powder form).

As clearly described, the present invention can provides aphysiologically useful substance for maintaining the human health fromresidue of saccharification of malt, which has been worthless andimproved value of the malt residue, a by- product of saccharification ofmalt. Furthermore, the obtained physiologically active extract can beused for the raw materials of functional foods and drinks or aneffective ingredient of medicines.

What is claimed is:
 1. A composition comprised of:1) An extract derivedfrom malt residue having the following properties:(i) said extract is afraction having a molecular weight from 19.8×10⁴ to 22.1×10⁴ daltons asdetermined by the GPC-LALLS method, (ii) Infrared absorption spectra at3,600-3,200 cm⁻¹ and 1,700-1,600 cm⁻¹, (iii) Proton nuclear magneticresonance spectra having peaks at 0.5-6.0 ppm and 6.0-8.5 ppm, (iv)Ultraviolet absorption spectra at 200-250 nm and 250-350 nm, (v)Positive to phenol sulfuric acid color reaction, (vi) Positive tocopper-Folin color reaction, and 2) an acceptable carrier or diluent. 2.A food which contains the composition of claim 1 as a food additive. 3.A beverage which contains the composition of claim 1 as a beverageadditive.
 4. The beverage of claim 2 which is milk.
 5. The beverage ofclaim 3 which is a powdered drink.
 6. The beverage of claim 3 which isorange juice.